title: Targeting transforming growth factor βRII expression inhibits the activation of hepatic stellate cells and reduces collagen synthesis.
abstract: Abnormal production of extracellular matrix (ECM) components significantly contributes to the development of liver fibrosis. This study aimed at examining the effects of short-hairpin RNA (shRNA)-mediated transient knockdown of transforming growth factor βRII (TGFβRII) expression on the proliferation and activation of hepatic stellate cells (HSCs) and synthesis of fibrogenic ECM components in HSC cells. Three different shRNA-expressing plasmids were constructed for the expression of shRNA-(a, b, c) targeting to the rat TGFβRII mRNA beginning at nucleotide position 339, 444 and 528 and they were transfected into a rat stellate cell line, HSC-T6 cells, respectively. The levels of TGFβRII, α-smooth muscle actin (α-SMA), and type I and III collagen expressions were characterized by reverse transcription polymerase chain reaction and Western blot assays. The concentrations of hyaluronic acid (HA) and type IV collagen in the supernatants of cultured cells were measured by enzyme-linked immunosorbent assay. Transfection with the TGFβRII-specific shRNAs resulted in varying levels of inhibition in the expression of TGFβRII in HSC-T6 cells, and transfection with the potent shRNA-c inhibited the expression of TGFβRII in a dose-dependent manner. Knockdown of TGFβRII expression significantly reduced the levels of α-SMA, type I, III and IV collagen, and HA expression in HSC-T6 cells (P < 0.01). In conclusion, our data indicated that knockdown of TGFβRII expression inhibited the activation of HSCs and the production of fibrogenic ECM components in HSC-T6 cells. Therefore, our findings support the notion that TGFβRII is an important factor of the pathogenic process of liver fibrosis.
authors.affiliation: Department of Infectious Diseases, the Third Affiliated Hospital of Wenzhou Medical College, Rui'an, Zhejiang, China.
authors.forename: Rongquan
authors.initials: R
authors.lastname: Fu
authors.forename: Jinguo
authors.initials: J
authors.lastname: Wu
authors.forename: Jiguang
authors.initials: J
authors.lastname: Ding
authors.forename: Jifang
authors.initials: J
authors.lastname: Sheng
authors.forename: Liang
authors.initials: L
authors.lastname: Hong
authors.forename: Qingfeng
authors.initials: Q
authors.lastname: Sun
authors.forename: Hui
authors.initials: H
authors.lastname: Fang
authors.forename: Dairong
authors.initials: D
authors.lastname: Xiang
mesh_terms: D000199:Actins / Q000096:biosynthesis; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation; D003094:Collagen / Q000096:biosynthesis*; D020869:Gene Expression Profiling; D055785:Gene Knockdown Techniques; D055166:Hepatic Stellate Cells / Q000378:metabolism / Q000502:physiology*; D006820:Hyaluronic Acid / Q000096:biosynthesis; D010957:Plasmids; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics; D051381:Rats; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000199:Actins; D003094:Collagen; D006820:Hyaluronic Acid; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta; C498862:Acta2 protein, rat